Cell biology

The research activities of this area at the cross-roads of materials science, nanotechnology and cell biology, primarily focusing on how newly-discovered materials and interfacial processes can be developed and used for specific applications. In particular, our research emphasizes cytomechanic studies correlating cytomechanical profiles to the underlying molecular and cellular mechanisms to isolate and study extracellular vesicles, with materials engineering promoting and driving cell adhesion and differentiation.

Cellular homeostasis and cytomechanics

The surface of living cells is strictly related to many cellular processes such as adhesion, signalling, transport, energy transformation, tumour metastasis etc. Cellular functions are mediated by a plethora of specific biomolecules including cell-to-cell adhesion proteins and those that constitute the highly complex and dynamic architecture of the cytoskeleton which, in turn, connects structurally and functionally the intracellular environments with the extracellular matrix via other adhesion molecules.

Our research activities focus on the biomolecular and cytomechanical cellular behaviour of primary fibroblast cultures related to juvenile hereditary Parkinson’s disease and of epithelial tumour cell lines. Nanomechanical properties of single cells obtained by advanced microscopic techniques allow to correlate morpho-mechanical cellular state changes upon interaction with pharmacological agents and the underlying biological \ biochemical processes.

From Left to right: Cover image of Ref. 4, selected publications; CLSM micrographs of organization of F-actin (stained with phalloidin, in red) and microtubules (stained with a-Tub, in green) filaments in fibroblasts cells (nuclei stained with DAPI, in blue) (Ref.3); Laser scanning confocal microscopy (left) and Scanning Force Microscopy image (right) of MCF-7 cells grown on Fn modified Petri dish. Cells were stained with TRITC-labelled phalloidin to reveal F-actin and with Hoechst to detect nuclei. When grown on Fn, the cells exhibited short actin-rich extensions around their perimeters, as indicated by arrows, as well as cytoplasmic extensions at the cell edges.

Analysis of Extracellular vesicles

A recently developed research activity concerns the EVs isolation from fibroblasts of parkin-mutant patients and the characterization of their biochemical differences compared to EV derived from control samples. In particular, EVs are isolated from culture media by an optimized differential centrifugation protocol and their size and morphology determined by flow cytometry, electron and atomic force microscopy. We are also performing a mass spectrometry analysis to investigate their lipidomic profiles.

Materials for cell engineering

Cell engineering is a very promising research field which aims to induce specific biological processes such as proliferation and differentiation, cell-to-cell interaction, biomolecular production and extracellular matrix (ECM) formation, at the cell/material interface. Specifically cell migration is a phenomenon that is involved in different physiological processes such as morphogenesis, wound healing and tumour invasion. Biochemical or biophysical stimuli such as chemotaxis, galvanotaxis extracellular matrix compliance/stiffness and topology can influence cell migration in terms of speed, direction and persistence. The control of the cell environment by multiple physicochemical cues has therefore emerged as a key factor to enable functionality, modulate response, and affect cell behaviour.

In particular our recent studies have shown that cells are able to recognize the mechanical properties of a substratum over which they move and that these properties direct the motion through a phenomenon called durotaxis. Thus, mechanical interactions between a cell and its underlying substratum play a crucial role in modulating cell motility. We are investigating combinations of external stimuli ranging from chemical to mechanical and electrical cues in the cellular microenvironment. Moreover we are carrying out cell tracking analysis to deepen our understanding of the mechanisms underlying cell motility.

The nature of the interface between cells and materials can stimulate a repulsive (or adhesive) response that can causes the cells to separate (adhere) to the desired material. This effect dramatically depends on surface properties of the material. Plasma surface modification is applicable whenever the surface of the device has to be bioactive or bioinert by changing chemical/topographical features of a material surface without affecting its bulk properties. As an example, the irreversible, undesired adhesion of biomolecules and cells (i.e. ‘biofouling’) can be controlled by a plasma assisted deposition of Polyethylene oxide (PEO)-like coatings. On the other hand, surfaces containing polar groups (i.e NH2, COOH, OH etc. …) or deposition of coatings in which biological molecules are dispersed in an organic matrix can be produced by plasma in order to impart bioactivity and biocompatibility to the surface. Finally, surfaces with micro- and nanostructured coatings can dramatically improve cell/material interactions due to topographical cues.

Sketch of the potentialities of plasma processing of surfaces in order to produce micro-nanostructured surfaces (left); unfouling surfaces (top), biomimetic/bioactive surfaces in which a coating embedding active biomolecules are deposited (right) and functional surfaces containing chemical groups like amino, carbonyl, alcoholic and carboxylic ones (bottom)

Plasma processing of cells and biological liquids

Cold atmospheric pressure plasmas are emerging as an exciting development for therapeutics. These plasmas are very efficient sources of highly reactive oxygen and nitrogen species (RONS), UV radiation, electromagnetic fields and charged particles. Experiments show that cold atmospheric plasmas allow efficient, contact-free and painless disinfection, without damaging healthy tissue. In healthcare, new horizons are being opened for wound healing, tissue regeneration, cancer therapy, and treatment of chronic wounds assisted by plasma technology.

Our facilities have been designed to allow us to study the response of different type of cells to different plasma doses and to correlate the chemical composition of plasma treated cell culture media with cell behaviour.

Abstract: Process for the production by plasmochemical deposition of a film having a nanometric thickness, optionally multilayered, permitting carrying out in a controlled, uniform and long lasting way, release of substances of interest in a surrounding medium containing liquids, from a substrate including the substance to be released as micro/nano particles, or from a layer deposited on the substrate including the substance to be released as micro/nano particles, or from a layer of the substance to be released deposited on the substrate, or from a substrate that is the substance to be released optionally in the form of particles. The substances to be released can be metals, compounds having anti-bacterial properties, biologically active molecules such as drugs, hormones, vegetable extracts, peptides, lipids, protides and glucides. The layer with the substance to be released, be it organic or inorganic, is obtained by plasmochemical deposition optionally having a structure similar to polyethylene oxide (PEO) or polyethylene glycol (PEG), called PEO-like polymers, constituted, in a variable percentage da ethylene oxide units (-CH2CH2O-, EO); barrier film is obtained by depositing by plasma at least one organic or inorganic layer, optionally with a PEO-like structure, wherein chemical composition, degree of crosslinking and thickness are adjustable by the plasmo chemical deposition process parameters, and allow to adjust the release of the active substance according to specific needs. The structures on which the above said films can be deposited are: medical-surgical devices, common handworks, structures known as scaffolds, and the above defined substances to be released themselves. The invention also relates to medical-surgical devices, common handworks and scaffolds coated by a substrate and barrier layer, as well as to biologically active substances coated by at least one barrier layer.

Project

My First AIRC Grant – Role of Electro/Mechanical cues in the control and guidance of Glioma Progression (MFAG) 2015 n. 16803 (2015-2018)

Latest News

The IV edition of TERAMETANANO, the International Conference on Terahertz Emission, Metamaterials and Nanophotonics, will take place in Lecce (Italy) from 27 to 31 of May 2019 in the 16th-century Castle of Charles V with two special nights that will be held in an original Theatre of Roman period.

TERAMETANANO is an annual conference that gather physicists studying a wide variety of phenomena in the areas of nano-structures, nano-photonics and meta-materials, with special attention to the coupling between light and matter and in a broad range of wavelengths, going from the visible up to the terahertz.

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